Effects of inflammation on developing glia

描述（由申请人提供）：脑瘫（CP）是影响成千上万儿童的最常见发育疾病之一。 CP是由大脑白色物质的正常发育障碍引起的。 妊娠妇女的子宫内炎症可能导致高达12%的CP病例。 动物模型的开发对于确定炎症和CP之间相关性的机制至关重要。 候选人已经开发了一种使用妊娠大鼠的子宫内炎症模型，该模型要么对发育中的少突胶质细胞造成损害，要么在发育中的大脑中显示出细胞因子反应。 据推测，细胞因子，释放的炎症刺激，是负责在宫内炎症过程中发育中的少突胶质细胞的损害。 候选人提出了一个为期五年的多学科研究和培训计划，重点是在两个相关的炎症实验模型中测试这一假设，包括转基因动物。 为此，他将确定实验性子宫内炎症对大鼠少突胶质细胞系细胞增殖和分化的影响。 候选人将在蛋白质和mRNA水平上表征胎儿大脑和胎盘中的细胞因子反应。 他将在小鼠中建立一个类似的炎症模型，并使用缺乏TNF受体的转基因动物来确定TNF-α在损伤发育中的少突胶质细胞中的作用。 他将使用一种转基因小鼠品系，在少突胶质细胞特异性启动子下游插入一个增强型绿色荧光蛋白的报告基因，在炎症刺激后通过荧光激活细胞分选（FACS）从这些细胞中纯化少突胶质细胞前体细胞。 通过提取RNA，他将比较实验刺激后参与炎症级联反应的基因表达。 该职业奖项将通过特定技术（RT-PCR、蛋白质印迹、使用基因微阵列的基因表达谱分析和报告标记细胞的流式细胞仪分选）的指导和培训来扩展候选人的能力。 此外，它将为他提供分析和解释从遗传到细胞水平解决炎症复杂病理生理过程所需数据的技能。 在神经科学导师以及智力迟钝和发育障碍研究中心的资源将支持这些明确的培训目标。

DESCRIPTION (provided by applicant): Cerebral palsy (CP) is one of the most common developmental diseases affecting thousands of children. CP is caused by disturbances of the normal development of the cerebral white matter. Intrauterine inflammation in pregnant women may be responsible for up to 12% of the cases of CP. Development of animal models is essential to determine the mechanisms responsible for the association between inflammation and CP. The candidate has developed a model of intrauterine inflammation using pregnant rats that either causes damage to developing oligodendrocytes or demonstrates a cytokine response in the developing brain. It is hypothesized that cytokines, released in response to the inflammatory stimulus, are responsible for damage to developing oligodendrocytes during intrauterine inflammation. The candidate proposes a five-year, multidisciplinary research and training program that focuses on testing this hypothesis in two related experimental models of inflammation, including transgenic animals. To this end, he will determine the effects of experimental intrauterine inflammation on the proliferation and differentiation of cells of the oligodendrocyte lineage in rats. The candidate will characterize the cytokine response in fetal brain and placenta at the protein and mRNA level. He will develop a similar inflammatory model in mice and use transgenic animals lacking TNF receptors to determine the role of TNF-( in the damage to developing oligodendrocytes. He will use a transgenic mouse strain that has a reporter gene for enhanced green fluorescent protein inserted downstream of an oligodendrocyte-specific promoter to purify oligodendrocyte precursor cells from these cells by fluorescence-activated cell sorting (FACS) after the inflammatory stimulus. By extracting RNA, he will compare expression of genes involved in the inflammatory cascade after the experimental stimulus. This career award will extend the candidate's capabilities through mentorship and training in specific techniques (RT-PCR, Western blot, gene expression profiling using gene microarrays and FACS sorting of reporter-labeled cells). In addition, it will provide him with skills in the analysis and interpretation of data required to address the complex pathophysiological processes of inflammation from the genetic to cellular level. Mentorship in neuroscience as well as the resources of the Mental Retardation and Developmental Disabilities Research Center will support these well-defined training objectives.